Figure 2 - available from: Journal of Inflammation
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Effect of hyaluronic acid on pro-inflammatory cytokine secretion. a Schematic drawing of the in vitro inflammatory co-culture setup of differentiated monolayer M1 macrophages on the top and human monolayer osteoarthritic (OA) chondrocytes at the bottom. b Quantification of pro-inflammatory cytokine concentration in culture medium in the presence or absence of hyaluronic acid
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Background
Osteoarthritis (OA) is described by an imbalance between anabolic and catabolic processes in the affected joint. This dysregulation of metabolism affects not only chondrocytes within cartilage tissue but also the cells of the synovial membrane across the border of the joint. An important factor in OA is the low viscosity of the synovial...
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... of hyaluronic acid (HA) as an anti- inflammatory agent in inflammatory diseases has been proposed and investigated in in vitro and in vivo condi- tions [20]. We intended to evaluate the role of high- molecular-weight (high-MW) HA on the inflammatory response on human osteoarthritic chondrocytes in our established in vitro co-culture system (Fig. 2a). Our re- sults have shown that incubation of OA chondrocytes with high-MW HA (1600 kDa) leads to a reduced con- centration of secreted IL-1ß and TNF-α cytokines into the medium, compared to the control group (Fig. 2b). We also observed a trend towards a lower concentration of secreted IL-6 for the HA-treated group, which how- ever did ...
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... (high-MW) HA on the inflammatory response on human osteoarthritic chondrocytes in our established in vitro co-culture system (Fig. 2a). Our re- sults have shown that incubation of OA chondrocytes with high-MW HA (1600 kDa) leads to a reduced con- centration of secreted IL-1ß and TNF-α cytokines into the medium, compared to the control group (Fig. 2b). We also observed a trend towards a lower concentration of secreted IL-6 for the HA-treated group, which how- ever did not reach statistical significance. The above results possibly suggest the anti-inflammatory properties of high-MW HA or a potential activation of anti- inflammatory mechanisms in human OA ...
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... system, with or without the addition of hyaluronic acid. b Comparative differentiation index between collagen type II and collagen type I in human osteoarthritic monolayer chondrocytes in the co-culture system, with or without the addition of hyaluronic acid in cultured human OA chondrocytes, present in our established transwell co-culture (Fig. 2a). Our results have shown a significantly reduced secretion of pro- inflammatory IL-1ß and TNF-α cytokines, but not IL-6 in the HA-treated group, compared to the control group (Fig. 2b). Also, high-MW HA treatment of human OA chondrocytes in our co-culture system resulted in an in- creased gene expression of anabolic cartilage markers ...
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... in the co-culture system, with or without the addition of hyaluronic acid in cultured human OA chondrocytes, present in our established transwell co-culture (Fig. 2a). Our results have shown a significantly reduced secretion of pro- inflammatory IL-1ß and TNF-α cytokines, but not IL-6 in the HA-treated group, compared to the control group (Fig. 2b). Also, high-MW HA treatment of human OA chondrocytes in our co-culture system resulted in an in- creased gene expression of anabolic cartilage markers (i.e. COL2A1 and ACAN) and reduced catabolic car- tilage markers (i.e. MMP13 and MMP3), compared to the control group (Fig. 3a). Chondrocytes tend to de- differentiate in prolonged 2D ...
Citations
... This study also demonstrated the ability of high-molecular-weight hyaluronic acid (HMWHA) to protect chondrocytes against the inflammatory response. 8 Collectively, these findings indicate the intricate communication and regulatory mechanisms that can be unveiled through coculture systems. ...
Osteoarthritis (OA) is a chronic musculoskeletal disorder characterized by cartilage degeneration and synovial inflammation. Paracrine interactions between chondrocytes and macrophages play an essential role in the onset and progression of OA. In this study, in replicating the inflammatory response during OA pathogenesis, chondrocytes were treated with interleukin-1β (IL-1β), and macrophages were treated with lipopolysaccharide and interferon-γ. In addition, a coculture system was developed to simulate the biological situation in the joint. In this study, we examined the impact of hyaluronic acid (HA) viscosupplement, particularly Hyruan Plus, on chondrocytes and macrophages. Notably, this viscosupplement has demonstrated promising outcomes in reducing inflammation; however, the underlying mechanism of action remains elusive. The viscosupplement attenuated inflammation, showing an inhibitory effect on nitric oxide production, downregulating proinflammatory cytokines such as matrix metalloproteinases (MMP13 and MMP3), and upregulating the expression levels of type II collagen and aggrecan in chondrocytes. HA also reduced the expression level of inflammatory cytokines such as IL-1β, TNF-α, and IL-6 in macrophages, and HA exerted an overall protective effect by partially suppressing the MAPK pathway in chondrocytes and p65/NF-κB signaling in macrophages. Therefore, HA shows potential as a viscosupplement for treating arthritic joints.
... The superior performance of the HA and Boswellia serrata extract supplements may be attributed to the synergistic effects of increased synovial HA concentration, which enhances joint lubrication and shock absorption [24], and the potent anti-inflammatory properties of Boswellia serrata [25]. In addition to contributing to the viscoelastic properties of synovial fluid, HA may also exhibit antiinflammatory and chondroprotective actions by modulating intra-and extracellular inflammation cascades [26], potentially contributing to its clinical efficacy in reducing joint pain and preserving cartilage health. Furthermore, a previous animal study demonstrated that administration of standardized Boswellia serrata extracts in a rat model of OA resulted in the suppression of inflammatory enzymes such as cyclooxygenase-2 and 5-lipoxygenase, leading to a significant reduction in prostaglandin E2 and leukotriene B4 levels [27]. ...
Background: Hyaluronic acid (HA), glucosamine (Glc), and chondroitin sulfate (CS) are key ingredients commonly incorporated into dietary chondroprotective supplements for the management of osteoarthritis (OA). Despite their widespread use, there is a paucity of published data regarding their efficacy and safety, necessitating rigorous investigation in clinical settings. To address this knowledge gap, we conducted a randomized, single-blind pilot study to evaluate the effects of two commercially available multi-ingredient supplements on patients with mild-to-moderate knee OA.
Methods: A total of 51 patients diagnosed with mild-to-moderate knee OA were enrolled in a four-week randomized study and allocated equally (1:1:1 ratio) into three groups: a control group (n = 17) that received no treatment, an HA group (n = 17) given Syalox® 300 Plus (1 tablet/day) containing HA (300 mg) and Boswellia serrata extract (100 mg), and a Glc + CS group (n = 17) given Cartijoint® Forte (1 tablet/day) containing Glc (415 mg), CS (400 mg), and curcuminoids from rhizomes of Curcuma longa L (50 mg).Physicians conducting evaluations were blinded to group assignments, whereas patients were not. All participants underwent assessments of pain relief, functional capacity improvement, and serum adropin levels, an emerging biomarker of knee OA, at baseline and after the four-week intervention period.
Results: Both the HA and the Glc + CS groups exhibited improvements at the end of the study relative to baseline, with statistically significant differences (p < 0.05) observed in pain at rest, pain during movement, range of motion, and the overall Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, including its pain, stiffness, and physical function subscales. Notably, the HA group outperformed the Glc + CS group in the alleviation of pain at rest, pain during movement, and on the WOMAC pain subscale, with all differences being statistically significant (p < 0.05). Additionally, both groups showed a significant elevation in serum adropin levels from baseline (p < 0.05), with the HA group experiencing a more substantial increase when compared to the Glc + CS group (p < 0.05). Both supplements showed a limited number of treatment-emergent adverse events.
Conclusion: Oral supplementation with either HA or Glc + CS demonstrated potential benefits for managing symptoms of mild-to-moderate knee OA. Notably, HA supplementation was associated with greater improvements in pain relief and higher elevations in serum adropin levels compared to Glc + CS supplementation. However, larger-scale and longer-term studies are necessary to further evaluate the safety and efficacy of these dietary supplements within the clinical management arsenal for knee OA.
... The utilization of HA in clinical practice for osteoarthritis treatment has previously evidenced its capacity to bolster the lubricating and shock-absorbing features of synovial fluid [16]. Additionally, HA can enact disease-modifying effects such as inflammation reduction [17] or pain relief, with concomitant safe use, even with multiple administrations [18]. GCs, despite their propensity to trigger chondrocyte apoptosis [19,20], are also applied in clinical practice for osteoarthritis treatment due to their anti-inflammatory effects. ...
... These observations are consistent with the literature [25,37] and suggest that the treatment induces conditions typically present in osteoarthritis. HA supplementation did not elicit any changes, even though high-molecular-weight HA typically exhibits anti-inflammatory effects [38,39], as previously demonstrated in one of our studies [17]. Adding GCs to cytokine treatment (GCs and GCs/HA) attenuated the negative impacts of inflammation, leading to increased expression of anabolic genes and reduced expression of catabolic genes. ...
Patients with knee osteoarthritis often receive glucocorticoid (GC) or hyaluronic acid (HA) injections to alleviate symptoms. This study evaluated the impact of Triamcinolone Hexacetonide (a GC), HA, and a combination of both on bovine osteochondral grafts exposed to IL-1β and IL-17 in an ex vivo culture. Metabolic activity increased with GC treatment. GCs and GCs/HA counteracted cytokine effects, with gene expressions similar to untreated controls, while HA alone did not. However, HA improved the coefficient of friction after two weeks. The highest friction values were observed in GC-containing and cytokine-treated groups. Cytokine treatment reduced tissue proteoglycan content, which HA could mitigate, especially in the GC/HA combination. This combo also effectively controlled proteoglycan release, supported by reduced sGAG release. Cytokine treatment led to surface cell death, while GCs, HA, or their combination showed protective effects against inflammation. The GC/HA combination had the best overall results, suggesting its potential as a superior treatment option for osteoarthritis.
... Обсуждение. Широко используемым вариантом лечения ОА являются в/с инъекции ГлК [26]. В суставах, пораженных ОА, концентрация ГлК в синовиальной жидкости всегда снижена по сравнению со здоровыми суставами [27]. ...
Osteoarthritis (OA) is the most common joint disease that affects more than 80% of people over 55 years and in its final stages leads to disability. One of the safe non-surgical methods of OA treatment is intra-articular injections of hyaluronic acid (HA).
Objective : to compare the viscoelastic rheological properties of HA medical products with different concentrations of sodium hyaluronate (SH) available in the Russian Federation.
Material and methods. The study was carried out using a modular rheometer MCR 302 (Anton Paar, Austria). All measurements were carried out at a temperature of 25.0±0.1 °C using the measuring system "cone-plane" (angle – 2 ˚ , cone diameter – 40 mm, gap height – 0.169 mm). The determination of the elastic moduli (G’) and viscosity (G”) was carried out depending on the frequency in the linear region of the shear stress, and the dynamic viscosity was determined at a shear rate of 1 sec-1.
Results. The evaluation of the rheological properties of the studied samples revealed a positive relationship with the concentration of SH and no relationship with the molecular weight of SH. The highest viscoelastic properties were possessed by medical products in the 1% SH group: Armaviscon and Ripart; in the group with SH concentration of 1.5–1.6%, all the studied samples, except for Hyalubrix, showed similar higher results compared to the previous group; in the group with SH concentration of 2–3%, Armaviscon Platinum had the highest results and Flexotron Ultra and Armaviscon Forte – somewhat lower results.
Conclusion. The study of the rheological viscoelastic properties of HA medical products is the most accessible method, on the basis of which it is possible to predict the clinical effect.
... Biomimetic nanoparticles and Lipid-based NPs are consisted form biocompatible components of cell membranes, whiteout induction of inflammatory neutrophil infiltration, and ROS production (Gary et al., 2022). High molecular weight (HMW) HA-coated ECMmimicking biomaterials elicit an anti-inflammatory role by promoting Treg (Bollyky et al., 2009) and reducing M1 macrophage differentiation (Bauer et al., 2016). In addition, nanomaterials-carrying immunomodulatory drugs present these potentials including (i) packaging of lower dosages of systemic drugs; (ii) delivery of drugs in localized, sustained, and controlled manners; (iii) providing a large functional surface for improved bioavailability and biodegradability; and (iv) delivery of typical drugs having poor solubility (Yao and Martins, 2020). ...
... OA is a chronic degenerative disease of the joints that leads to pain and loss of mobility [59]. Intra-articular injection of high molecular weight HA to replace synovial fluid that has lost its viscoelastic properties is widely used in the treatment for OA [60]. HA plays a dual role in the process of inflammation and damage of cartilage, as a proinflammatory molecule or an anti-inflammatory molecule [31] It basically depends on the molecular weight of HA. ...
High molecular weight hyaluronic acids (HMW-HAs) have been used for the palliative treatment of osteoarthritis (OA) for decades, but the pharmacological activity of HA fragments has not been fully explored due to the limited availability of structurally defined HA fragments. In this study, we synthesized a series glycosides of oligosaccharides of HA (o-HAs), hereinafter collectively referred to as o-HA derivatives. Their effects on OA progression were examined in a chondrocyte inflammatory model established by the lipopolysaccharide (LPS)-challenged ATDC5 cells. CCK-8 assays and RT-qPCR showed that o-HA derivatives (≤100 μg/mL) exhibited no cytotoxicity and inflammatory effects. We found that the o-HAs derivatives alleviated LPS-induced inflammation, apoptosis, autophagy and proliferation-inhibition of ATDC5 cells, similar to the activities of HMW-HAs. Moreover, Western blot analysis showed that different HA derivatives selectively reversed the effects of LPS on the expression of extracellular matrix (ECM)-related proteins (MMP13, COL2A1 and Aggrecan) in ATDC5 cells. Our study suggested that o-HA derivatives may alleviate LPS-induced chondrocyte injury by reducing the inflammatory response, maintaining cell proliferation, inhibiting apoptosis and autophagy, and decreasing ECM degradation, supporting a potential oligosaccharides-mediated therapy for OA.
... For cocultivation, a transwell (Corning, USA) coculture system was employed as described previously [32,33]. Briefly, the differentiated M0 macrophages and the polarized M1 macrophages as described above were seeded in the upper chamber, and the HFL-1 cells were plated into the bottom chamber for cocultivation for 48 h. ...
Idiopathic pulmonary fibrosis (IPF) is a chronic progressive interstitial lung disease that leads rapidly to death. The present study is aimed at discovering the in-depth pathogenesis of IPF, exploring the role of adiponectin/carnitine palmityl transferase 1A- (APN/CPT1A-) mediated fatty acid metabolism during the development of IPF, and excavating its potential mechanism. Here, THP-1 cells were differentiated into M0 macrophages, followed by polarization to M1 macrophages upon hypoxia. Subsequently, lung fibroblast HFL-1 cells were stimulated by M1 macrophages to simulate hypoxia-related IPF condition in vitro. It was discovered that the stimulation of M1 macrophages promoted fibroblast proliferation and fibrosis formation in vitro, accompanied with a disorder of the APN/CPT1A pathway, an overproduction of lipid peroxides, and a low level of autophagy in HFL-1 cells. Thereafter, APN treatment or CPT1A overexpression greatly suppressed above lipid peroxide accumulation, fibroblast proliferation, and fibrosis but activated autophagy in vitro. Furthermore, an in vivo IPF rat model was established by injection of bleomycin (BLM). Consistently, CPT1A overexpression exerted a protective role against pulmonary fibrosis in vivo; however, the antifibrosis property of CPT1A was partly abolished by 3-methyladenine (an autophagy inhibitor). In summary, APN/CPT1A-mediated fatty acid metabolism exerted its protective role in IPF partly through activating autophagy, shedding a new prospective for the treatment of IPF.
... An important role in infrapatellar fat pad inflammation and fibrosis has also recently been discovered [3]. Today, the treatment modalities for OA include non-pharmacological (e.g., physiotherapy), pharmacological (e.g., steroidal and nonsteroidal anti-inflammatory drugs), or intra-articular (e.g., injection of hyaluronic acid) therapies [4,5]. These classical therapies can reverse the symptoms only in a small number of cases, but they do not stop the degeneration process of the cartilage or promote the repair of the tissue. ...
Background:
In recent decades, hyaluronic acid (HA) has attracted great attention as a new treatment option for osteoarthritis. Classical therapies are not able to stop the cartilage degeneration process nor do they favor tissue repair. Nowadays, it is accepted that high molecular weight HA can reduce inflammation by promoting tissue regeneration; therefore, the aim of this study was to verify the efficacy of a new high molecular weight HA of plant origin (called GreenIuronic®) in maintaining joint homeostasis and preventing the harmful processes of osteoarthritis.
Methods:
The bioavailability of GreenIuronic® was investigated in a 3D intestinal barrier model that mimics human oral intake while excluding damage to the intestinal barrier. Furthermore, the chemical significance and biological properties of GreenIuronic® were investigated in conditions that simulate osteoarthritis.
Results:
Our data demonstrated that GreenIuronic® crosses the intestinal barrier without side effects as it has a chemical-biological profile, which could be responsible for many specific chondrocyte functions. Furthermore, in the osteoarthritis model, GreenIuronic® can modulate the molecular mechanism responsible for preventing and restoring the degradation of cartilage.
Conclusion:
According to our results, this new form of HA appears to be well absorbed and distributed to chondrocytes, preserving their biological activities. Therefore, the oral administration of GreenIuronic® in humans can be considered a valid strategy to obtain beneficial therapeutic effects during osteoarthritis.
... In addition, the combination product showed increased sGAG synthesis [30], which could not be shown in our study. The results indicate that high-molecular-weight HA at the used concentration is constructive to alleviate the cytokine-induced proteoglycan catabolism and matrix turnover, as previously reported in a study [31]. ...
... Adding test substances to the cytokine-treated chondrocytes should reverse the negative effect on gene expression patterns, as high-molecular-weight HA and GC have anti-inflammatory effects [33]. The former has already been shown in a co-culture study with HA from another manufacturer [31]. In the present study, there was a non-significant difference in the cartilage-specific genes COL2A1, ACAN and SOX9. ...
Intra-articular injections of glucocorticoids (GC) or hyaluronic acid (HA) are commonly used interventions for patients suffering from knee osteoarthritis (OA). Both substances are combined to achieve a chondroprotective and anti-inflammatory effect. Clinical studies have shown benefits, but data on the cellular level are still lacking. This study aimed to investigate the effect of the GC triamcinolone hexacetonide, HA, and a mix of both substances on cytokine-treated chondrocytes in vitro. Chondrocytes isolated from human articular cartilage were seeded on 6- and 24-well plates. Mimicking OA’s inflammatory state, cells were treated with IL-1β and IL-17 for six days, whereby, after three days, test substances (10%) were added to the culture medium. Chondrocytes were analyzed on days three and six concerning their actin polymerization, expression of anabolic and catabolic genes, metabolic activity, cytokine release, and reactive oxygen species (ROS). Adding HA or GC/HA to the inflammatory culture medium increased the metabolic activity of chondrocytes, while groups containing GC reduced catabolic gene expression and the release of TNF-α. In addition, enhanced F-actin content was shown supplementing HA or GC/HA to the culture medium. Supplementing GC with HA leads to an anti-inflammatory and chondroprotective effect by diminishing the side effects of GC supplementation alone.
... High-MW HA has been shown to promote M1-to-M2 phenotype switch, which results in dampening of the production of pro-inflammatory molecules (including MMPs) and stimulation of anti-inflammatory molecules [64][65][66]. Also, HMW HA represses the production of MIF in some cells [67] and the production of inflammatory cytokines/MMPs in other cells (e.g., chondrocytes) [68][69][70]. These observations could explain our finding of reduced M1 macrophages within HA-immersed CM discs, either by direct effects of HA on macrophages or through neighboring, host cells. ...
Objectives
We previously showed that accelerated degradation of collagen membranes (CMs) in diabetic rats is associated with increased infiltration of macrophages and blood vessels. Since pre-implantation immersion of CMs in cross-linked high molecular weight hyaluronic acid (CLHA) delays membrane degradation, we evaluated here its effect on the number of macrophages and endothelial cells (ECs) within the CM as a possible mechanism for inhibition of CM resorption.
Materials and methods
Diabetes was induced with streptozotocin in 16 rats, while 16 healthy rats served as control. CM discs were labeled with biotin, soaked in CLHA or PBS, and implanted under the scalp. Fourteen days later, CMs were embedded in paraffin and the number of macrophages and ECs within the CMs was determined using antibodies against CD68 and transglutaminase II, respectively.
Results
Diabetes increased the number of macrophages and ECs within the CMs (∼2.5-fold and fourfold, respectively). Immersion of CMs in CLHA statistically significantly reduced the number of macrophages (p < 0.0001) in diabetic rats, but not that of ECs. In the healthy group, CLHA had no significant effect on the number of either cells. Higher residual collagen area and membrane thickness in CLHA-treated CMs in diabetic animals were significantly correlated with reduced number of macrophages but not ECs.
Conclusions
Immersion of CM in CLHA inhibits macrophage infiltration and reduces CM degradation in diabetic animals.
Clinical relevance
The combination of CLHA and CM may represent a valuable approach when guided tissue regeneration or guided bone regeneration procedures are performed in diabetic patients.
